Abstract

Aim 1 is to test the hypothesis that alternative initiation motifs for helix 12 regulate conformational changes in ER, and hence regulate function.
Aim 2 will test the hypothesis that Asp 351 interacts with helix 12 and thereby stabilizes the active conformation.
Aim 3 will test the hypothesis that the antagonist activity of two modified estrogens is due to interference with proper initiation of helix 12 and/or interference with the interaction between Asp 351 and helix 12. The proposed research will use mutational analysis, ligand binding studies, circular dichroism spectroscopy, as well as measurements of protein stability, transactivation ability, and association with coactivators and corepressors.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK)
Type
Research Project (R01)
Project #
5R01DK056934-03
Application #
6524463
Study Section
Biochemical Endocrinology Study Section (BCE)
Program Officer
Margolis, Ronald N
Project Start
2000-09-15
Project End
2004-08-31
Budget Start
2002-09-01
Budget End
2004-08-31
Support Year
3
Fiscal Year
2002
Total Cost
$186,250
Indirect Cost

  Institution

Name
Wayne State University
Department
Physiology
Type
Schools of Medicine
DUNS #
City
Detroit
State
MI
Country
United States
Zip Code
48202

  Publications

Koide, Akiko; Zhao, Changqing; Naganuma, Misuzu et al. (2007) Identification of regions within the F domain of the human estrogen receptor alpha that are important for modulating transactivation and protein-protein interactions. Mol Endocrinol 21:829-42
Rossetti, Sandro; Consugar, Mark B; Chapman, Arlene B et al. (2007) Comprehensive molecular diagnostics in autosomal dominant polycystic kidney disease. J Am Soc Nephrol 18:2143-60
Harris, Peter C; Bae, Kyongtae T; Rossetti, Sandro et al. (2006) Cyst number but not the rate of cystic growth is associated with the mutated gene in autosomal dominant polycystic kidney disease. J Am Soc Nephrol 17:3013-9
Zhao, Changqing; Abrams, Judith; Skafar, Debra F (2006) Targeted mutation of key residues at the start of helix 12 in the hERalpha ligand-binding domain identifies the role of hydrogen-bonding and hydrophobic interactions in the activity of the protein. J Steroid Biochem Mol Biol 98:1-11
Zhao, Changqing; Koide, Akiko; Abrams, Judith et al. (2003) Mutation of Leu-536 in human estrogen receptor-alpha alters the coupling between ligand binding, transcription activation, and receptor conformation. J Biol Chem 278:27278-86
Schwartz, Janice A; Zhong, Li; Deighton-Collins, Sarah et al. (2002) Mutations targeted to a predicted helix in the extreme carboxyl-terminal region of the human estrogen receptor-alpha alter its response to estradiol and 4-hydroxytamoxifen. J Biol Chem 277:13202-9
Zhong, L; Skafar, D F (2002) Mutations of tyrosine 537 in the human estrogen receptor-alpha selectively alter the receptor's affinity for estradiol and the kinetics of the interaction. Biochemistry 41:4209-17